US2016116448A1PendingUtilityA1
pH-MODULATED QUENCHERS OF FLUORESCENCE
Est. expirySep 30, 2034(~8.2 yrs left)· nominal 20-yr term from priority
G01N 21/643C07C 255/57C07D 213/76G01N 2021/6432G01N 31/221C07C 317/44G01N 2021/6439C07C 205/56
33
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Claims
Abstract
The present disclosure relates to a compound of formula (I) or a salt thereof, wherein variable A, B, and X are as defined herein. The disclosure also relates to covalent conjugates of formula (II). The disclosure further relates to the use of the compounds of formula (I) and covalent conjugates of formula (II) in methods for detecting a change in the pH of a mixture.
Claims
exact text as granted — not AI-modified1 . A compound of formula (I)
or a salt thereof, wherein:
A is CO 2 R 1 , CO 2 R 2 , C(O)NR 3 R 4 , CN, S(O)R 5 , SO 2 R 5 , C(O)R 6 , C(O)R 7 , or CO(NHS);
B is selected from the group consisting of B 1 , B 2 , B 3 , and B 4 ; wherein
B 1 is
B 2 is selected from the group consisting of
B 3 is selected from the group consisting of
and
B 4 is selected from the group consisting of
each X is independently N, C—H, C—Z 1 , or C—Z 2 ;
each Q is independently H, NO 2 , Cl, Br, F, I, CH 3 , OCH 3 , CN, CO 2 R 9 , C(O)NR 10 R 11 , SO 2 R 12 , SOR 12 , CF 3 or N 3 ;
R 1 is H or (C 1 -C 12 )alkyl;
R 2 is CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, (CH 2 ) m OH, (CH 2 ) m SH, (CH 2 ) m N 3 , (CH 2 ) m NH 2 , (CH 2 ) m CO(NHS), (CH 2 ) m (N-maleimide), (CH 2 ) m O(CEP), (CH 2 ) m OCH 2 C≡CH, (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, (CH 2 CH 2 O) n CH 2 CH 2 OH, (CH 2 CH 2 O) n CH 2 CH 2 SH, (CH 2 CH 2 O) n CH 2 CH 2 N 3 , (CH 2 CH 2 O) n CH 2 CH 2 NH 2 , (CH 2 CH 2 O) n CH 2 CH 2 CO(NHS), (CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide), (CH 2 CH 2 O) n CH 2 CH 2 O(CEP), (CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 OH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT) or (CH 2 CH 2 ) n OCH 2 CH(O(CEP))CH 2 O(DMT);
R 3 is H, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, (CH 2 ) m OH, (CH 2 ) m SH, (CH 2 ) m N 3 , (CH 2 ) m NH 2 , (CH 2 ) m CO(NHS), (CH 2 ) m (N-maleimide), (CH 2 ) m O(CEP), (CH 2 ) m OCH 2 C≡CH, (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, (CH 2 CH 2 O) n CH 2 CH 2 OH, (CH 2 CH 2 O) n CH 2 CH 2 SH, (CH 2 CH 2 O) n CH 2 CH 2 N 3 , (CH 2 CH 2 O) n CH 2 CH 2 NH 2 , (CH 2 CH 2 O) n CH 2 CH 2 CO(NHS), (CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide), (CH 2 CH 2 O) n CH 2 CH 2 O(CEP), (CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 OH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT) or (CH 2 CH 2 ) n OCH 2 CH(O(CEP))CH 2 O(DMT);
R 4 is H, (C 1 -C 6 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, or R 3 and R 4 together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
R 5 is (C 1 -C 6 )alkyl or phenyl, wherein said phenyl may be unsubstituted or substituted with up to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
R 6 is H, (C 1 -C 6 )alkyl, or phenyl;
R 7 is phenyl, wherein said phenyl is substituted with one to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
R 8 is H, (C 1 -C 6 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 9 is H, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 10 is H, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 11 is H, (C 1 -C 6 )alkyl, or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, or R 10 and R 11 together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
R 12 is (C 1 -C 6 )alkyl or phenyl, wherein said phenyl may be unsubstituted or substituted with up to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
each Y is independently N, C—H, C—Z 1 , or C—Z 2 ;
each Z 1 is independently OCH 3 , CN, or CF 3 ;
each Z 2 is independently CH 3 , F, Cl, Br, or I;
m is an integer from 2 to 8; and
each n is independently an integer from 1 to 5;
wherein if each X is independently C—H or C—Z 2 , and A is CO 2 R 1 , CN, or COR E , then B is B 2 , B 3 , or B 4 .
2 . The compound or salt of claim 1 , wherein each
X is C—H, A is CO 2 R 1 , and B is B 2 , B 3 , or B 4 ; each X is C—H, A is CN, and B is B 2 , B 3 , or B 4 ; each X is C—H, A is COR 6 , and B is B 2 , B 3 , or B 4 ; at least one X is N or C—Z 1 and A is CO 2 R 1 , CN or COR 6 ; or A is CO 2 R 2 , C(O)NR 3 R 4 , S(O)R 5 , SO 2 R 5 , C(O)R 7 , or CO(NHS).
3 . (canceled)
4 . (canceled)
5 . (canceled)
6 . (canceled)
7 . (canceled)
8 . (canceled)
9 . (canceled)
10 . The compound or salt of claim 1 , wherein A is CO 2 R 1 and R 1 is (C 1 -C 12 )alkyl;
A is CO 2 R 2 and R 2 is CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl; A is CO 2 R 2 and R 2 is (CH 2 ) OH, (CH 2 ) m SH, (CH 2 ) m NH 2 , (CH) m CO(NHS) or —(CH 2 ) m (N-maleimide); A is CO 2 R 2 and R 2 is —(CH 2 CH 2 O) n CH 2 CH 2 OH, —(CH 2 CH 2 O) n CH 2 CH 2 SH, —(CH 2 CH 2 O) n CH 2 CH 2 NH 2 , —(CH 2 CH 2 O) n CH 2 CH 2 CO(NHS) or —(CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide); A is CO 2 R 2 and R 2 is —(CH 2 ) m N 3 , —(CH 2 CH 2 O)CH 2 CH 2 N 3 , —(CH 2 ) m OCH 2 C≡CH or —(CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH; A is CO 2 R 2 and R 2 is —(CH 2 ) m O(CEP), —(CH 2 CH 2 O) n CH 2 CH 2 O(CEP), —(CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, —(CH 2 CH 2 O) n CH 2 CH(OH)CH 2 OH, —CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), —(CH 2 CH 2 O) n CH 2 CH(OH)CH 2 O(DMT), —(CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT) or —(CH 2 CH 2 O) n CH 2 CH(O(CEP))CH 2 O(DMT); A is C(O)NR 3 R 4 and R 3 is (C 1 -C 12 )alkyl; A is C(O)NR 3 R 4 and R 3 is (CH 2 ) m OH, (CH 2 ) m SH, (CH 2 ) m NH 2 , (CH 2 ) m CO(NHS) or —(CH 2 ) m (N-maleimide); A is C(O)NR 3 R 4 and R 3 is —(CH 2 CH 2 O) n CH 2 CH 2 OH, —(CH 2 CH 2 O) n CH 2 CH 2 SH, —(CH 2 CH 2 O) n CH 2 CH 2 NH 2 , —(CH 2 CH 2 O) n CH 2 CH 2 CO(NHS) or —(CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide); A is C(O)NR 3 R 4 and R 3 is (CH 2 ) m N 3 , —(CH 2 CH 2 O) n CH 2 CH 2 N 3 , —(CH 2 ) m OCH 2 C≡CH or —(CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH; A is C(O)NR 3 R 4 and R 3 is —(CH 2 ) m O(CEP), —(CH 2 CH 2 O) n CH 2 CH 2 O(CEP), —(CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, —(CH 2 CH 2 O) n CH 2 CH(OH)CH 2 OH, —(CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), —(CH 2 CH 2 O) n CH 2 CH(OH)CH 2 O(DMT), —(CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT) or —(CH 2 CH 2 O) n CH 2 CH(O(CEP))CH 2 O(DMT); or A is C(O)NR 3 R 4 and R 3 and R 4 together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
11 . (canceled)
12 . (canceled)
13 . (canceled)
14 . (canceled)
15 . (canceled)
16 . (canceled)
17 . (canceled)
18 . (canceled)
19 . (canceled)
20 . (canceled)
21 . (canceled)
22 . The compound or salt of claim 1 , wherein the compound of formula (I) is a compound of formula (Ia)
a compound of formula (Ib)
a compound of formula (Ic)
a compound of formula (Id)
a compound of formula (Ie)
a compound of formula (If)
or
a compound of formula (Ig)
23 . (canceled)
24 . (canceled)
25 . (canceled)
26 . (canceled)
27 . (canceled)
28 . (canceled)
29 . The compound or salt of claim 1 , wherein each X is C—H.
30 . The compound or salt of claim 1 , wherein each Y is C—H.
31 . The compound or salt of claim 1 , wherein at least one occurrence of X is N.
32 . The compound or salt of claim 1 , wherein at least one occurrence of Y is N.
33 . (canceled)
34 . (canceled)
35 . (canceled)
36 . (canceled)
37 . The compound or salt of claim 1 , wherein the compound or salt is selected from the group consisting of:
Hexyl 2,2-bis(2,4-dinitrophenyl)acetate;
Isopropyl 2-(2,4-dinitrophenyl)-2-(3,5-dinitropyridin-2-yl)acetate;
2-(2,4-dinitrophenyl)-2-(3,5-dinitropyridin-2-yl)-N, N-diethylacetamide;
3-(3-(bis(4-methoxyphenyl)(phenyl)methoxy)-2-hydroxypropoxy)propyl 2-(2,4-dinitrophenyl)-2-(3,5-dinitropyridin-2-yl)acetate;
2-(2,4-dicyanophenyl)-2-(3,5-dinitropyridin-2-yl)-N,N-diethylacetamide;
2-(3,5-dinitropyridin-2-yl)-N,N-diethyl-2-(4-(phenylsulfonyl)phenyl)acetamide;
2-(2,4-dinitrophenyl)-2-(3,5-dinitropyridin-2-yl)-1-(piperidin-1-yl)ethan-1-one;
2-(3,5-dinitropyridin-2-yl)-N,N-diethyl-2-(4-(methylsulfonyl)phenyl)acetamide;
2-(2,4-dinitrophenyl)-N,N-diethyl-2-(4-(phenylsulfonyl)phenyl)acetamide; and
2,5-dioxopyrrolidin-1-yl 1-(2-(2,4-dinitrophenyl)-2-(3,5-dinitropyridin-2-yl)acetyl)piperidine-4-carboxylate;
or a salt thereof.
38 . The compound or salt of claim 1 , wherein the compound or salt is:
Hexyl 2-(2-cyano-4-nitrophenyl)-2-(2,4-dinitrophenyl)acetate;
or a salt thereof.
39 . The compound or salt of claim 1 , wherein:
A is CO 2 R 1 , CO 2 R 2 , C(O)NR 3 R 4 , CN, S(O)R 5 , SO 2 R 5 , C(O)R 6 , or C(O)R 7 ; R 2 is CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl; R 3 is H, (C 1 -C 12 )alkyl, or CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl; and R 4 is H, (C 1 -C 6 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, or R 3 and R 4 together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
40 . A covalent conjugate of formula (II)
or a salt thereof, wherein,
A′ is CO 2 R 1 , CO 2 R 2′ , C(O)NR 3′ R 4′ , CN, S(O)R 5 , SO 2 R 5 , C(O)R 6 , C(O)R 7 , CO(NHS), or CO-(L-[Conjugated Species]);
B is selected from the group consisting of B 1 , B 2 , B 3 , and B 4 ; wherein
B 1 is
B 2 is selected from the group consisting of
B 3 is selected from the group consisting of
and
B 4 is selected from the croup consisting of
each X is independently N, C—H, C—Z 1 , or C—Z 2 ;
L is a single covalent bond or a covalent linkage having 1-50 non-hydrogen atoms selected from the group consisting of C, N, O, S and P and is composed of any combination of single, double, triple or aromatic bonds;
each Q′ is independently H, NO 2 , Cl, Br, F, I, CH 3 , OCH 3 , CN, CO 2 R 9 , C(O)NR 1′ R 11′ , SO 2 R 12 , SOR 12 , CF 3 , N 3 , or CO-(L-[Conjugated Species]);
R 1 is H or (C 1 -C 12 )alkyl;
R 2′ is CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, (CH 2 ) m OH, (CH 2 ) m SH, (CH 2 ) m N 3 , (CH 2 ) m NH 2 , (CH 2 ) m CO(NHS), (CH 2 ) m (N-maleimide), (CH 2 ) m O(CEP), (CH 2 ) m OCH 2 C≡CH, (CH 2 ) m -L-[Conjugated Species], (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, (CH 2 CH 2 O) n CH 2 CH 2 OH, (CH 2 CH 2 O) n CH 2 CH 2 SH, (CH 2 CH 2 O) n CH 2 CH 2 N 3 , (CH 2 CH 2 O) n CH 2 CH 2 NH 2 , (CH 2 CH 2 O) n CH 2 CH 2 CO(NHS), (CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide), (CH 2 CH 2 O) n CH 2 CH 2 O(CEP), (CH 2 CH 2 O) n CH 2 CH 2 -L-[Conjugated Species], (CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH, (CH 2 CH 2 O) n CH 2 -L-[Conjugated Species], (CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(L-[Conjugated Species])CH 2 OH, (CH 2 ) 3 OCH 2 CH(L-[Conjugated Species])CH 2 -L-[Conjugated Species], (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 OH, (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 O(DMT), (CH 2 CH 2 ) n OCH 2 CH(O(CEP))CH 2 O(DMT), (CH 2 CH 2 ) n OCH 2 CH(L-[Conjugated Species])CH 2 OH, or (CH 2 CH 2 ) n OCH 2 CH(L-[Conjugated Species])CH 2 -L-[Conjugated Species];
R 3′ is H, (C 1 -C 12 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, (CH 2 ) m OH, (CH 2 ) m SH, (CH 2 ) m N 3 , (CH 2 ) m NH 2 , (CH 2 ) m CO(NHS), (CH 2 ) m (N-maleimide), (CH 2 ) m O(CEP), (CH 2 ) m OCH 2 C≡CH, (CH 2 ) m -L-[Conjugated Species], (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, (CH 2 CH 2 O) n CH 2 CH 2 OH, (CH 2 CH 2 O) n CH 2 CH 2 SH, (CH 2 CH 2 O) n CH 2 CH 2 N 3 , (CH 2 CH 2 O) n CH 2 CH 2 NH 2 , (CH 2 CH 2 O) n CH 2 CH 2 CO(NHS), (CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide), (CH 2 CH 2 O) n CH 2 CH 2 O(CEP), (CH 2 CH 2 O) n CH 2 CH 2 -L-[Conjugated Species], (CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH, (CH 2 CH 2 O) n CH 2 -L-[Conjugated Species], (CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(L-[Conjugated Species])CH 2 OH, (CH 2 ) 3 OCH 2 CH(L-[Conjugated Species])CH 2 -L-[Conjugated Species], (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 OH, (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 O(DMT), (CH 2 CH 2 ) n OCH 2 CH(O(CEP))CH 2 O(DMT), (CH 2 CH 2 O) n OCH 2 CH(L-[Conjugated Species])CH 2 OH, or (CH 2 CH 2 O) n OCH 2 CH(L-[Conjugated Species])CH 2 -L-[Conjugated Species];
R 4′ is H, (C 1 -C 6 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, or R 3′ and R 4′ together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
R 5 is (C 1 -C 6 )alkyl or phenyl, wherein said phenyl may be unsubstituted or substituted with up to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
R 6 is H, (C 1 -C 6 )alkyl, or phenyl;
R 7 is phenyl, wherein said phenyl is substituted with one to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
Fe is H, (C 1 -C 6 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 9 is H, (C 1 -C 12 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 10′ is H, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 11′ is H, (C 1 -C 6 )alkyl, or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, or R 10′ and R 11′ together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
R 12 is (C 1 -C 6 )alkyl or phenyl, wherein said phenyl may be unsubstituted or substituted with up to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
each Y is independently N, C—H, C—Z 1 , or C—Z 2 ;
each Z 1 is independently OCH 3 , CN, or CF 3 ;
each Z 2 is independently CH 3 , F, Cl, Br, or I;
m is an integer from 2 to 8; and
each n is independently an integer from 1 to 5;
wherein the covalent conjugate of formula (II) includes at least one occurrence of -L-[Conjugated Species].
41 . The covalent conjugate of claim 40 , wherein each X is C—H.
42 . The covalent conjugate of claim 40 , wherein each Y is C—H.
43 . The covalent conjugate of claim 40 , wherein at least one occurrence of X is N.
44 . The covalent conjugate of claim 40 , wherein at least one occurrence of Y is N.
45 . (canceled)
46 . (canceled)
47 . (canceled)
48 . (canceled)
49 . The covalent conjugate or salt of any one of claim 40 , wherein the Conjugated Species is a protein, an antibody, a peptide, a nucleic acid, an oligonucleotide, a solid support, a soluble polymer, an insoluble polymer, a dendrimer, a saccharide, a fatty acid, a lipid, or a phospholipid.
50 . (canceled)
51 . (canceled)
52 . (canceled)
53 . (canceled)
54 . (canceled)
55 . (canceled)
56 . A method for detecting a change in the pH of a mixture, comprising:
providing a mixture comprising a fluorescence donor and the compound or salt of claim 1 ; irradiating the resulting mixture at a first time with light having a wavelength suitable to excite the fluorescence donor; measuring detectable fluorescence emitted by the fluorescence donor while irradiating the mixture at said first time; irradiating the resulting mixture at a second time with light having a wavelength suitable to excite the fluorescence donor; measuring detectable fluorescence emitted by the fluorescence donor while irradiating the mixture at said second time; and comparing the detectable fluorescence emitted by the fluorescence donor at said first time with the detectable fluorescence emitted by the fluorescence donor at said second time to detect a change in the pH of the mixture; wherein a decrease in detectable fluorescence at said second time relative to said first time indicates an increase in the pH of the mixture, and wherein an increase in detectable fluorescence at said second time relative to said first time indicates a decrease in the pH of the mixture.
57 . A method for detecting a change in the pH of a mixture, comprising:
providing a mixture comprising a fluorescence donor and a compound of formula (I)
or a salt thereof, wherein:
A is CO 2 R 1 , CO 2 R 2 , C(O)NR 3 R 4 , CN, S(O)R 5 , SO 2 R 5 , C(O)R 6 , C(O)R 7 , or CO(NHS);
B is selected from the group consisting of B 1 , B 2 , B 3 , and B 4 ; wherein
B 1 is
B 2 is selected from the group consisting of
B 3 is selected from the group consisting of
and
B 4 is selected from the group consisting of
each X is independently N, C—H, C—Z 1 , or C—Z 2 ;
each Q is independently H, NO 2 , Cl, Br, F, I, CH 3 , OCH 3 , CN, CO 2 R 9 , C(O)NR 10 R 11 , SO 2 R 12 , SOR 12 , CF 3 or N 3 ;
R 1 is H or (C 1 -C 12 )alkyl;
R 2 is CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, (CH 2 ) m OH, (CH 2 ) m SH, (CH 2 ) m N 3 , (CH 2 ) m OCH 2 C≡CH, (CH 2 ) m NH 2 , (CH 2 ) m CO(NHS), (CH 2 ) m (N-maleimide), (CH 2 ) m O(CEP), (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, (CH 2 CH 2 O) n CH 2 CH 2 OH, (CH 2 CH 2 O) n CH 2 CH 2 SH, (CH 2 CH 2 O) n CH 2 CH 2 N 3 , (CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH, (CH 2 CH 2 O) n CH 2 CH 2 NH 2 , (CH 2 CH 2 O) n CH 2 CH 2 CO(NHS), (CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide), (CH 2 CH 2 O) n CH 2 CH 2 O(CEP), (CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 OH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT) or (CH 2 CH 2 ) n OCH 2 CH(O(CEP))CH 2 O(DMT);
R 3 is H, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, (CH 2 ) m OH, (CH 2 ) m SH, (CH 2 ) m N 3 , (CH 2 ) m OCH 2 C≡CH, (CH 2 ) m NH 2 , (CH 2 ) m CO(NHS), (CH 2 ) m (N-maleimide), (CH 2 ) m O(CEP), (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, (CH 2 CH 2 O) n CH 2 CH 2 OH, (CH 2 CH 2 O) n CH 2 CH 2 SH, (CH 2 CH 2 O) n CH 2 CH 2 N 3 , (CH 2 CH 2 O) n CH 2 CH 2 OCH 2 C≡CH, (CH 2 CH 2 O) n CH 2 CH 2 NH 2 , (CH 2 CH 2 O) n CH 2 CH 2 CO(NHS), (CH 2 CH 2 O) n CH 2 CH 2 (N-maleimide), (CH 2 CH 2 O) n CH 2 CH 2 O(CEP), (CH 2 ) 3 OCH 2 CH(OH)CH 2 OH, (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 OH, (CH 2 ) 3 OCH 2 CH(OH)CH 2 O(DMT), (CH 2 CH 2 ) n OCH 2 CH(OH)CH 2 O(DMT), (CH 2 ) 3 OCH 2 CH(O(CEP))CH 2 O(DMT) or (CH 2 CH 2 ) n OCH 2 CH(O(CEP))CH 2 O(DMT);
R 4 is H, (C 1 -C 6 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl, or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, or R 3 and R 4 together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
R 5 is (C 1 -C 6 )alkyl or phenyl, wherein said phenyl may be unsubstituted or substituted with up to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
R 6 is H, (C 1 -C 6 )alkyl, or phenyl;
R 7 is phenyl, wherein said phenyl is substituted with one to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
R 8 is H, (C 1 -C 6 )alkyl, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 9 is H, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 10 is H, CH 2 CH 2 (C 4 -C 12 )perfluoroalkyl), or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl;
R 11 is H, (C 1 -C 6 )alkyl, or (CH 2 CH 2 O) n CH 2 CH 2 O(C 1 -C 4 )alkyl, or R 10 and R 11 together with the nitrogen atom to which they are attached form a ring selected from the group consisting of
R 12 is (C 1 -C 6 )alkyl or phenyl, wherein said phenyl may be unsubstituted or substituted with up to five substituents selected from the group consisting of CH 3 , F, Cl, Br, I, OCH 3 , OH, NO 2 , CN, CF 3 , N 3 , and N(CH 3 ) 2 ;
each Y is independently N, C—H, C—Z 1 , or C—Z 2 ;
each Z 1 is independently OCH 3 , CN, or CF 3 ;
each Z 2 is independently CH 3 , F, Cl, Br, or I;
m is an integer from 2 to 8;
and
each n is independently an integer from 1 to 5;
irradiating the resulting mixture at a first time with light having a wavelength suitable to excite the fluorescence donor;
measuring detectable fluorescence emitted by the fluorescence donor while irradiating the mixture at said first time;
irradiating the resulting mixture at a second time with light having a wavelength suitable to excite the fluorescence donor; and
measuring detectable fluorescence emitted by the fluorescence donor while irradiating the mixture at said second time; and
comparing the detectable fluorescence emitted by the fluorescence donor at said first time with the detectable fluorescence emitted by the fluorescence donor at said second time to detect a change in the pH of the mixture;
wherein a decrease in detectable fluorescence at said second time relative to said first time indicates an increase in the pH of the mixture, and wherein an increase in detectable fluorescence at said second time relative to said first time indicates a decrease in the pH of the mixture.
58 . A method for detecting a change in the pH of a mixture, comprising:
providing a mixture comprising a fluorescence donor and the covalent conjugate or salt of claim 40 ; irradiating the resulting mixture at a first time with light having a wavelength suitable to excite the fluorescence donor; measuring detectable fluorescence emitted by the fluorescence donor while irradiating the mixture at said first time; irradiating the resulting mixture at a second time with light having a wavelength suitable to excite the fluorescence donor; measuring detectable fluorescence emitted by the fluorescence donor while irradiating the mixture at said second time; and comparing the detectable fluorescence emitted by the fluorescence donor at said first time with the detectable fluorescence emitted by the fluorescence donor at said second time to detect a change in the pH of the mixture; wherein a decrease in detectable fluorescence at said second time relative to said first time indicates an increase in the pH of the mixture, and wherein an increase in detectable fluorescence at said second time relative to said first time indicates a decrease in the pH of the mixture.Join the waitlist — get patent alerts
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